Calvaria lysate WT1
Total RNA was extracted from isolated osteoblasts using the Qiagen RNeasy Micro Extraction Kit (Qiagen, UK, cat no: 74004) and RNA integrity and quantity analysed with High Sensitivity RNA ScreenTape (Agilent, California) and purity determined using the Qubit High Sensitivity RNA assay kit (ThermoFisher Scientific, UK, cat no: Q10210). 50 ng RNA per sample was used to generate libraries using the Lexogen QuantSeq 3 mRNA-Seq Library Prep Kit FWD for Illumina (Lexogen, US). Equal amounts (4 nM per sample) were sequenced: v2.5 150 cycles Mid output (75 single read) flowcell, NextSeq 500 (Illumina, California).